The minimal bactericidal concentrations

(MBC) were expressed as the range a–b, in which a corresponds to the highest concentration in which bacterial growth was observed and b corresponds to the lowest concentration that kills GSK126 order 100% of the cells. Three biological replicates were used for the determination of MBC. Total RNA was isolated from mid-log suspensions of the strain 9a5c incubated or not with 50 μM of gomesin at 28 °C for 15, 30 and 60 min using Trizol reagent (Invitrogen, Carlsbad, CA). The RNA concentration was determined using an ND-1000 spectrophotometer (Thermo Scientific, Wilmington, DE) after treatment with RQ1 RNAse-free DNAse (Promega, Madison, WI), and its reliability was evaluated by electrophoresis on formaldehyde-agarose gels. cDNA labeling and hybridization of microarray slides were performed as described previously (Zaini et al., 2008). A detailed description of the microarray can be found in Koide et al. (2004) and Zaini et al. (2008) and at the NCBI’s Gene Trichostatin A molecular weight Expression Omnibus (GEO) database (http://www.ncbi.nlm.nih.gov/geo) under accession number GPL2708. Data represent six biological replicates with at least two technical replicates each. Microarray data acquisition, normalization and analysis were performed as detailed by Koide et al. (2004). A gene was considered differentially expressed when >66.5% of the biological replicates were

outside the intensity-dependent cutoff curves obtained by self–self hybridization experiments (Koide et al., 2004; Zaini et al., 2008) and exhibited a fold-change >2.0. The complete data set has been submitted to the GEO database according to MIAME guidelines and is accessible through GEO series accession number GSE17605 (http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE17605).

Pyruvate dehydrogenase lipoamide kinase isozyme 1 RT-qPCR was performed in the GeneAmp 5700 thermocycler (Applied Biosystems, Carlsbad, CA) as described previously (Zaini et al., 2008). Data from two biological replicates with three technical replicates were used to calculate the relative changes in coding sequence (CDS) expression levels as described (Zaini et al., 2008). Mid-log suspensions of the strain 9a5c of X. fastidiosa were transferred to glass flasks and incubated with 50 μM gomesin, streptomycin 1 μg mL−1 or water as control. After 4 days at 28 °C, the walls of the glass flasks were examined to evaluate biofilm production as described (Kjaergaard et al., 2000). Two independent biological experiments were assayed in triplicate. Mid-log suspensions of strain 9a5c or strain J1a12 of X. fastidiosa were incubated with 25 or 50 μM gomesin or water as a control. After 18 h at 28 °C, cells were harvested by centrifugation at 3000 g for 10 min at 25 °C, washed twice in sterile phosphate-buffered saline (PBS) and suspended in PBS. The resulting bacterial suspensions were used to mechanically inoculate Nicotiana clevelandii plants as detailed (Lopes et al., 2000).

This conclusion aligns with that reached by Hughes et al.[42] when they evaluated Y 27632 the level of pharmaceutical care provided by community pharmacists within 13 European countries using the Behavioral Pharmaceutical Care Scale of pharmaceutical care in community pharmacies. The relative lack of patient-care-related terms and the fact that ‘medicine’ and ‘dispense’ were the most frequently reported terms indicate that

medicines rather than the patients are the main current focus of pharmacists when they consider their role.[43] Rosenthal et al.[24] reported that pharmacists’ reluctance to become more involved in patient-centred care provision can be explained by certain passive pharmacists’ characteristics, such as not having enough confidence in themselves, fear of taking risks and waiting for physicians’ approval. The findings of the present study suggest that product-focused

practice still predominates within the pharmacy profession in both Alberta and Northern Ireland. This may be explained by the fact that the pharmacists’ mental model, which is an internal image on the way the pharmacy profession works which prevents the pharmacist from thinking or acting in a different way,[25] still links pharmacy profession to product-focused practice. While the findings of the present study helped to explore certain aspects of current pharmacy culture in Northern Ireland and Alberta, there is a need for further exploration into pharmacy culture. A better understanding of the current pharmacy culture will help to use improved progression strategy see more to move the pharmacy profession into patient-centredness. Pharmacy culture must align with the desired changes, if a transition in pharmacy practice to a more patient-centred approach is to take place.[27] Community pharmacists

in Northern Ireland provided more patient-centred responses when compared to community pharmacists in Alberta. This could be explained by the fact that community pharmacists in Northern Ireland are paid to provide certain patient-centred services, such as minor ailments management and 4-Aminobutyrate aminotransferase smoking cessation. This can lead to the conclusion that community pharmacists may offer patient-centred services if they were offered sustainable remuneration. The relative lack of patient-care-related terms suggests that when it comes to the pharmacists’ practice in both Alberta and Northern Ireland patient care is still not their first priority. The findings of the present study suggest that product-focused practice still predominates within the pharmacy profession in both Alberta and Northern Ireland. The Author(s) declare(s) that they have no conflicts of interest to disclose. This research received no specific grant from any funding agency in the public, commercial, or not-for-profits sectors.

The mean intensity was calculated for each spot, Ai=log2(RiGi)0.5 (Dudoit et al., 2002). A normalization method based on local regression was applied according to Yang et al. (2002), Mi=log2(Ri/Gi)log2(Ri/Gi)−c(A)=log2(Ri/[kj(A)Gi]), where c(A) is the LOWESS (locally weighted scatter plot smoothing) fit to the MA plot. Significant up- or downregulation of genes was identified Lumacaftor cell line by t statistics (Dudoit et al., 2002). Genes were accounted as differentially expressed if P≤0.05 and M≥1.00 or ≤−1.00. Normalization and t statistics were

carried out using the emma 2.2 microarray data analysis software developed at the Bioinformatics Resource Facility, Center for Biotechnology, Bielefeld University (Dondrup et al., 2003). For scanning electron microscopy (SEM), cells were grown on Permanox slides in ONR7a with either 1.5% hexadecane or 2% pyruvate as the carbon/energy source. SEM was carried out as described by Lünsdorf et al. (2001). The microarray experiments were performed with the exponentially grown cells grown on either hexadecane or pyruvate (as control conditions), and led to the identification of a total of 220 differentially expressed genes, with 109 genes being upregulated and 111 genes being downregulated. Differentially expressed genes could be grouped into 15 functional

categories, according to designated this website metabolic functions of the corresponding gene products. Both upregulated and downregulated genes were found in most groups, Protirelin with the exception of those genes grouped under ‘alkane oxidation’, ‘stress’, and ‘iron uptake’, whose functions were exclusively induced in the presence of alkanes. ‘Nitrogen assimilation’ genes were

all found to be expressed on pyruvate only, as were a number of other genes known to enable the cells to assimilate essential macroelements other than N, namely phosphorus and sulfur from less favorable sources. This effect may at least partially be attributed to higher cell densities present in pyruvate cultures, leading to some scarcity of these macroelements in the pyruvate, but not yet in the alkane-grown cultures. In the following, we therefore focus primarily on the functions that were found to be upregulated on alkanes, and thus can most clearly be attributed to A. borkumensis responses to growth on alkanes. The presence of an enzymatic system mediating the terminal oxidation of alkanes distinguishes an alkane-degrader from a non-alkane-degrading organism. Our earlier proteomic study has already revealed the presence of several alternative ways for the terminal oxidation of alkanes by A. borkumensis (Sabirova et al., 2006). In accordance with the proteomic data, here, we find alkane monooxygenase alkB1 (ABO_2707, Table 1) to be upregulated on hexadecane. Moreover, a second alkane monooxygenase alkB2 (ABO_0122, Table 1) was also found to be upregulated, which corresponds to data using earlier reverse transcriptase-PCR (Schneiker et al.

The mean intensity was calculated for each spot, Ai=log2(RiGi)0.5 (Dudoit et al., 2002). A normalization method based on local regression was applied according to Yang et al. (2002), Mi=log2(Ri/Gi)log2(Ri/Gi)−c(A)=log2(Ri/[kj(A)Gi]), where c(A) is the LOWESS (locally weighted scatter plot smoothing) fit to the MA plot. Significant up- or downregulation of genes was identified this website by t statistics (Dudoit et al., 2002). Genes were accounted as differentially expressed if P≤0.05 and M≥1.00 or ≤−1.00. Normalization and t statistics were

carried out using the emma 2.2 microarray data analysis software developed at the Bioinformatics Resource Facility, Center for Biotechnology, Bielefeld University (Dondrup et al., 2003). For scanning electron microscopy (SEM), cells were grown on Permanox slides in ONR7a with either 1.5% hexadecane or 2% pyruvate as the carbon/energy source. SEM was carried out as described by Lünsdorf et al. (2001). The microarray experiments were performed with the exponentially grown cells grown on either hexadecane or pyruvate (as control conditions), and led to the identification of a total of 220 differentially expressed genes, with 109 genes being upregulated and 111 genes being downregulated. Differentially expressed genes could be grouped into 15 functional

categories, according to designated buy Galunisertib metabolic functions of the corresponding gene products. Both upregulated and downregulated genes were found in most groups, Fossariinae with the exception of those genes grouped under ‘alkane oxidation’, ‘stress’, and ‘iron uptake’, whose functions were exclusively induced in the presence of alkanes. ‘Nitrogen assimilation’ genes were

all found to be expressed on pyruvate only, as were a number of other genes known to enable the cells to assimilate essential macroelements other than N, namely phosphorus and sulfur from less favorable sources. This effect may at least partially be attributed to higher cell densities present in pyruvate cultures, leading to some scarcity of these macroelements in the pyruvate, but not yet in the alkane-grown cultures. In the following, we therefore focus primarily on the functions that were found to be upregulated on alkanes, and thus can most clearly be attributed to A. borkumensis responses to growth on alkanes. The presence of an enzymatic system mediating the terminal oxidation of alkanes distinguishes an alkane-degrader from a non-alkane-degrading organism. Our earlier proteomic study has already revealed the presence of several alternative ways for the terminal oxidation of alkanes by A. borkumensis (Sabirova et al., 2006). In accordance with the proteomic data, here, we find alkane monooxygenase alkB1 (ABO_2707, Table 1) to be upregulated on hexadecane. Moreover, a second alkane monooxygenase alkB2 (ABO_0122, Table 1) was also found to be upregulated, which corresponds to data using earlier reverse transcriptase-PCR (Schneiker et al.

The D10 values were compared by mean of the selleck Student t-test (P≤0.01). The nodulation kinetics of S. meliloti 2011 harvested from continuous cultures established at pH 7.0 and at pH 6.1 were analyzed

in plastic pouches with modified nitrogen-free Fåhraeus medium (Lodeiro et al., 2000) at both pH 7.0 (20 mM PIPES) and pH 5.6 (20 mM MES). Seeds of M. sativa cv. Monarca, INTA, Argentina, were surface-sterilized for 10 min with 30% v/v commercial bleach (equivalent to 55 g L−1 active Cl2), followed by six washes with sterile distilled water. The seeds were then germinated on 1.5% w/v water-agar. Two-day-old seedlings were transferred to ethylene oxide-sterilized plastic growth pouches containing 10 mL of the corresponding Fåhraeus solution. Three days later, each root was inoculated with c. 105 CFU of the corresponding rhizobia by dripping 100 μL of bacterial suspension onto the root from the tip to the base. The plants were cultured in a growth chamber at 22 °C, with a photoperiod of 16 h/8 h – day/night. The number and relative location – on the primary and secondary roots – of individual nodules were then scored for the 3 weeks that followed the inoculation. Plastic pots with 70 g of sterile vermiculite were used in the competition experiments. Five 2-day-old seedlings were transferred to each plastic pot irrigated with modified nitrogen-free Fåhraeus mineral

solution buffered check details at either pH 7.0 or 5.6. Five days later the alfalfa plants were inoculated with 50 mL of a mix containing 1 × 105 CFU mL−1 of S. meliloti 20MP6 (GFP) and 1 × 105 CFU mL−1 of S. meliloti 2011; each of them either acid-adapted (ATR+, grown in batch cuture at pH 6.1) or control Isotretinoin (ATR−, grown in batch culture at pH 7.0), as indicated. Thirty days after the inoculation, the root nodules were excised, washed, and observed under a Leica MZ8 fluorescence stereomicroscope (Leica Microsystems, Wetzlar, Germany) to detect the presence of strain S. meliloti 20MP6 within the nodules. Previous work from our laboratory had shown

that nodule co-occupancy with two inoculated genotypes represented an event of low incidence (Lagares et al., 1992). The number of nodules occupied by each rhizobia were log-normalized and analyzed by means of the Student t-test (P≤0.05). As indicated above, the bacterial adaptive ATR has been largely reported in enteric bacteria as well as in rhizobia. In order to investigate the conditions that are necessary for the generation of an ATR in S. meliloti, rhizobia were grown in batch-culture systems and in a chemostat under continuous cultivation, both at neutrality and under moderate acidity (pH=6.1), and the death rates of the resulting bacteria were evaluated at pH 4.0 (see Materials and methods). As previously shown for S. medicae WSM419 grown at pH 5.

In all the phosphorylation assays, samples were analysed by SDS-PAGE and autoradiography overnight. All 1D 1H NMR spectra were recorded at a 1H frequency of 700 MHz

on a Bruker Advance III spectrometer at 25 °C in a buffer containing 20 mM sodium phosphate, pH 8.0, and 150 mM NaCl using protein samples at 0.1 mM concentration. Bioinformatic analysis of a DNA sequence upstream of the arsenite oxidase gene aroB allowed for the identification of two ORFs (Fig. 1a). The first ORF, designated aroR, contains 1323 base Obeticholic Acid pairs encoding a putative protein of 441 amino acids; the second ORF, named aroS, contains 1470 base pairs encoding a putative protein of 490 amino acids. Analysis of AroS and AroR amino acid sequences revealed their

similarity to a typical two-component system signalling protein, where aroS codes for a sensor histidine kinase while aroR codes for a response regulator (Fig. 1b). The AroS protein is characterized Akt inhibitor by the presence of a dimerization and histidine phosphotransfer domain (DHp; residues 263–329) and an ATP-binding catalytic domain (CA; residues 370–480) in its C-terminus (Fig. 1b); the two domains are commonly found in a classical input component of a two-component signalling pathway. The DHp domain contains a conserved histidine residue that undergoes ATP-dependent phosphorylation, through the activity of the CA domain, in response to changes in the external environment. Sequence alignments identified the histidine residue located at position 273 as the presumed site of autophosphylation (Fig. 2a). In addition, AroS is predicted to contain two transmembrane segments within its N-terminus. Transmembrane segment 1 is proposed to include residues

14 through 32, while transmembrane segment 2 Staurosporine purchase lies between residues 175 and 194. Present between these two transmembrane segments is the environmental stimuli-sensing portion of the protein, the sensory domain. Sequence analysis of this domain revealed that although the NT-26 AroS protein shares significant sequence identity with sensory domains from soil bacteria A. tumefaciens (80%) and O. tritici (79%), no significant homologue of a known structure could be identified. However, the length of the domain, secondary structure prediction and a weak homology to other unrelated sensory proteins would suggest that the regions fold most likely into a PAS-like topology. Interestingly, no cysteine residues are present in NT-26 AroS, implying that arsenite sensing and binding does not involve thiolate, as it is the case in other known arsenite-binding proteins (Mizumura et al., 2010). In contrast the AroS homologue in A. tumefaciens does contain a Cys at position 401, which has been implicated in binding arsenite (Kashyap et al., 2006). Sequence analysis of AroR identified a canonical two-component response regulator receiver domain (residues 6–118) in the N-terminal region of the protein sequence (Fig. 1b).

Twenty transtibial amputees (16 male) aged 60.1 years (range

45–80 years), and 20 age- and gender-matched healthy adult controls were recruited. Single-pulse transcranial magnetic stimulation assessed corticomotor excitability. Two indices of corticomotor excitability were calculated. An index of corticospinal excitability (ICE) determined relative excitability of ipsilateral and contralateral corticomotor projections to alpha-motoneurons innervating the quadriceps muscle (QM) of MLN0128 solubility dmso the amputated limb. A laterality index (LI) assessed relative excitability of contralateral projections from each hemisphere. Spatial-temporal gait analysis was performed to calculate step-time variability. Amputees had lower ICE values, indicating relatively greater excitability of ipsilateral corticomotor Maraviroc mouse projections than controls (P = 0.04). A lower ICE value was associated with increased step-time variability for amputated (P = 0.04) and non-amputated limbs (P = 0.02). This association suggests corticomotor projections

from ipsilateral M1 to alpha-motoneurons innervating the amputated limb QM may interfere with gait. Cortical excitability in amputees was not increased bilaterally, contrary to our hypothesis. There was no difference in excitability of contralateral M1 between amputees and controls (P = 0.10), and no difference in LI (P = 0.71). It appears both hemispheres control one QM, with predominance of contralateral corticomotor excitability in healthy adults. Following lower-limb amputation, putative ipsilateral corticomotor excitability is relatively increased in some amputees and may negatively impact on function. “
“The lack of axonal regeneration in the adult central nervous system is in part attributable to the presence of inhibitory molecules present in the environment of injured axons such as the myelin-associated proteins Nogo-A and MAG and the repulsive guidance molecules Ephrins, Netrins and Semaphorins. In the present study, we hypothesized that EphA4 and one

of its potential binding partners EphrinA3 may participate in the inhibition of adult axon regeneration in the model of adult mouse optic nerve injury. Axonal regeneration Selleck Rucaparib was analysed in three dimensions after tissue clearing of EphA4 knockout (KO), EphrinA3 KO and wild-type (WT) optic nerves. By immunohistochemistry, EphA4 was highly expressed in Müller glia endfeet in the retina and in astrocytes in the retina and the optic nerve, while EphrinA3 was present in retinal ganglion cells and oligodendrocytes. Optic nerve crush did not cause expression changes. Significantly more axons grew in the crushed optic nerve of EphA4 KO mice than in WT or EphrinA3 KO animals. Single axon analysis revealed that EphA4 KO axons were less prone to form aberrant branching than axons in the other mouse groups.

, 2006; Nakashima et al., 2011). Consequently, RND-type drug transporters have been termed ‘cis transporters’. This is in contrast to drug transporters from the major facilitator superfamily and ATP-binding cassette families where drug transport occurs through an alternating access mechanism across the cytoplasmic membrane, the so-called trans-transporters (Eicher et al., 2009; Nikaido & Takatsuka, 2009; Doshi et al., 2011). However, studies on purified and reconstituted AcrD from E. coli revealed that this RND

transporter could efflux the CTLA-4 antibody inhibitor membrane-impermeable aminoglycoside gentamycin from either side of the liposomes, consistent with the existence of both periplasmic and cytoplasmic drug efflux pathways (Aires & Nikaido, 2005). It is therefore highly likely that, in addition to the periplasmic drug efflux pathway, a pathway exists for removal of drugs directly from the cytosol or inner membrane leaflet. Das and co-workers

(Das et al., 2007) predicted that conserved Phe residues on the small N-terminal helix of AcrB which line a 15 Å opening on the cytoplasmic side of AcrB could play a role in the discrimination, uptake and transport of drug molecules from the cytoplasm through the central cavity formed by the membrane domains of the AcrB trimer. Phenylalanine residues are hugely important in VE 822 substrate recognition and binding in the RND-type drug transporters. The binding protomer in the asymmetric AcrB structure forms a hydrophobic pocket lined by phenylalanines 136, 178, 610, 615, 617 and 628 (Murakami et al., 2006; Seeger et al., 2006; Sennhauser et al., 2009) while minocycline, rifampicin and erythromycin are in direct contact with Phe residues (Murakami et al., 2006; Nakashima et al., 2011). Replacement of these phenylalanines with alanines reduced the ability of AcrB to confer resistance to a wide range of compounds (Bohnert et al., 2008; Vargiu et al.,

2011), with the F610A mutation having the greatest effect. Similarly, site-directed mutagenesis of F386, F388, F458 and F459 in AcrB resulted in a decrease in the minimum inhibitory concentration (MIC) for tetracycline, erythromycin, dequalinium and acriflavine (Yu et al., 2005). In this study, we aimed to provide more insight on the role of the conserved phenylalanine residues on the small Cell Penetrating Peptide N-terminal helix of MexB in drug efflux. These Phe residues were mutated to Ala residues to generate FAFA MexB. The ability of the mutant protein to confer resistance to toxic compounds and to efflux dyes was compared to that of the wild-type protein. For cytotoxicity and transport assays, plasmids were propagated in E. coli strain BW25113 lacking either AcrB or AcrA and AcrB (a kind gift from Professor Martin Pos, Institute of Biochemistry, Goethe-University, Frankfurt am Main, Germany). The plasmids used were pET41a (+), pUC18 (Novagen) and derivatives expressing MexB with a C-terminal His tag (pMexBH; Barrera et al.

There is a need to improve the quality of reporting of mixed-methods research in pharmacy practice. The framework proposed in this article can ensure quality reporting of mixed-methods studies. Mixed-methods approaches have huge potential to develop, inform and improve the fast-growing discipline of pharmacy practice. The Authors declare that they have no conflicts of interest to disclose. This research received

no specific grant RG7422 cost from any funding agency in the public, commercial or not-for-profit sectors. MAH is receiving a 3-year PhD scholarship from School of Healthcare, University of Leeds. All Authors state that they had complete access to the study data that support the publication. All Authors contributed substantially in the development of the article and all Authors have read the final version of the article and approved it for submission. “
“The impact of patient aggression on healthcare staff has been an important research topic over the past decade. However, the majority of that research has focused primarily on hospital staff, with only a minority GDC 0199 of studies examining staff in primary care settings such as pharmacies or doctors’ surgeries. Moreover, whilst there is an indication that patient aggression can impact the quality of patient care, no research has been conducted to examine how the impact of aggression

on staff could affect patient safety. The aim of the current study was to examine the impact of aggression on community pharmacists in Scotland. Three main aspects were examined: the cause of patient aggression, the impact of aggression on pharmacist job performance and pharmacist behaviours in response to aggression. A sample of 18 community pharmacists were interviewed using the critical incident technique. In total, 37 incidents involving aggressive patients were transcribed. Aggression was considered by the majority of participants to be based on a lack ifenprodil of understanding about the role of a pharmacist. More worrying were the reports of near misses and dispensing errors occurring after an aggressive incident

had taken place, indicating an adverse effect on patient safety. Pharmacists described using non-technical skills, including leadership, task management, situational awareness and decision-making, in response to aggressive behaviour. Patient aggression may have a significant impact on patient safety. This could be addressed through training in non-technical skills but further research is required to clarify those skills in pharmacy staff. “
“Objective  Cardiovascular disease is a major public health problem despite established treatment guidelines and significant healthcare expenditure worldwide. Poor medication compliance accounts in part for some of the observed evidence/practice gaps. Trials of fixed-dose combination pills are currently underway, but the attitudes of relevant health professionals to the routine use of a cardiovascular polypill are generally unknown.

coli isolates from diseased piglets in Guangdong Province, China. It also describes the association between AMR and VGs, and between resistance and phylogenetic background. Other such studies describing associations between resistance and virulence traits have invariably investigated a limited number of antimicrobials (principally ampicillin, tetracycline, chloramphenicol, streptomycin, and sulfonamides), whereas we have extended our observations to include doxycycline, florfenicol, apramycin, and amikacin. Such studies, reporting an association between the resistance of this range of antimicrobials and VGs among E. coli strains from diseased swine in South China, are not available at

present.

The results from this study showed alarming frequencies of resistance to many antimicrobial agents CHIR-99021 solubility dmso commonly used in China. In agreement with previous reports (White et al., 2000; Lanz et al., 2003; Maynard et al., 2003), most E. coli isolates from swine were resistant to sulfamethoxazole, tetracycline, streptomycin, and chloramphenicol. Multidrug resistance phenotypes of E. coli isolates from animals have been reported worldwide (Lanz et al., 2003; Maynard et al., 2003; Yang et al., 2004), and in accordance with this, >50% of E. coli strains in our study were resistant to 8–10 antimicrobials tested. Doxcycline and florfenicol have been approved Akt inhibitor for use in food-producing animals in China, and are now used Ureohydrolase extensively with livestock, resulting in the emergence of resistance to both drugs. Many E. coli isolates showed high resistance or reduced susceptibility to doxycycline as well as to florfenicol in this study, which is similar to previous studies (Bischoff et al., 2002; Dai et al., 2008). The likely reasons for the high resistance rates are the inappropriate use of these antimicrobials in veterinary practice and cross-resistance among antibiotics of the same class, such as tetracycline and chloramphenicol, although chloramphenicol has been prohibited for use in food animals in China. Similarly high resistance rates to ciprofloxacin seen in this study have

also been observed in other studies in China among E. coli isolates from swine and chickens (Yang et al., 2004; Liu et al., 2007; Dai et al., 2008), which suggests that this agent has become ineffective in veterinary medicine in China (Xu, 2001). Ceftiofur is the only cephalosporin approved for systemic use in food-producing animals since 2002 in China, and it is highly effective against E. coli isolates. The rate of resistance to ceftiofur was higher in our study than in previous studies (Yang et al., 2004; Liu et al., 2007), presumably as a consequence of the increasing use of cephalosporins on animal farms. Prudent use of antimicrobials in veterinary practice is therefore fundamental to the reduction of resistance development.